This study aimed to get ready leaf extract-loaded chitosan nanoparticles (nano-ASLE) against human colon cancer (WiDr) cells
This study aimed to get ready leaf extract-loaded chitosan nanoparticles (nano-ASLE) against human colon cancer (WiDr) cells. flavonoids, alkaloids, saponins, tannins, glycosides, phenolic, terpenoids, and steroids. has been reported in the discovery of new Annonaceous acetogenins, called as (2,4-cis and trans)-squamolinone, (2,4-cis and trans)-9 oxoasimicinone, and bullacin B. Acetogenins have been isolated from the fruit pulp, seeds, twigs, roots, stems, leaves, and bark of many plants in the family. Acetogenins isolated from seeds reported cytotoxic activity through inhibited proliferation on HL-60 cells and induced apoptosis by the HSL-IN-1 activation Rabbit Polyclonal to Akt of caspase-3 and also against 9KB nasopharynx cells, A549 lung cells, and HT-29 colon cells. In the past, studies showed that green biosynthesis of silver nanoparticles of leaf extract was reported to have cytotoxic effect and induction of HSL-IN-1 apoptosis on MCF-7 cells. Nanotechnology is a promising approach to enhance the bioavailability of herbal medicine. Nontoxic biopolymer material is one of the nanoparticles which are expected to protect bioactive compound. A carrier material like the combination of chitosan and sodium tripolyphosphate (Na-TPP) is used to protect, stabilize, and control the release of the core. Chitosan can easily cross-link with Na-TPP as polyanions to control the release of the drug. Therefore, this study designed to prepare nano-A. Squamosa leaf extract (ASLE) HSL-IN-1 to enhance their bioactivities as an anticancer agent on WiDr cells due to induced apoptosis and cell cycle arrest. MATERIALS AND METHODS Ethical clearance All treatment procedures have been tested through The Medical and Health Research Ethics Committee, Faculty of Medicine, Gadjah Mada University, Yogyakarta, Indonesia (Approval Reference Number: KE/FK/0106/EC/2018). Preparation of leaf extract leaves were gathered from Lumajang Regency, East Java, Indonesia. The seed material was discovered at Purwodadi Botanical Backyard, Indonesian Institute of Sciences, Pasuruan, Indonesia, and accepted with the guide amount 003/IPH.06.HM/VII/2017. The leaves had been cleansed and cut into little parts and tone dried. They were powdered using mechanical blender and exceeded through the coarse sieve (0.2 mm). The powder was macerated with ethanol 96% for 72 h at 37C. The extract was evaporated in the water bath at the heat of 60C. The residue was then stored in a refrigerator at the heat of 0C4C. Preparation of leaf extract-loaded chitosan nanoparticles leaf extract-loaded chitosan nanoparticles (nano-ASLE) were prepared by ionic gelation of chitosan (0.1% w/v) and sodium tripolyphosphate (0.84% w/v) anions. One gram of was dissolved in 50 mL distilled water and added 100 mL chitosan in glacial acetic (0.25% v/v) and added 350 mL sodium tripolyphosphate in dropwise into the solution stirring condition at room temperature. The pH answer was adjusted by adding 0.1 M NaOH treatment for the chitosan-complex and stirred for 2 h on magnetic stirrer. Chitosan-was centrifuged at 6000 rpm for 30 min and decanted. The supernatant was collected and dried for onward usage as nano-ASLE. Particle size analysis Dynamic light-scattering method was performed to measure the particle size distribution of nano-ASLE using Zetasizer Nano ZS (Malvern Instruments Ltd., UK). Cell culture of WiDr cells WiDr cells were collected from your Parasitology Laboratory, Faculty of Medicine, Gadjah Mada University or college, Yogyakarta, Indonesia. Cells were cultured in DMEM and supplemented with 10% (v/v) fetal bovine serum, 3% streptomycin-penicillin, and 1% fungizone in 5% CO2 incubator at 37C. Determination of IC50value The IC50 value of the nano-ASLE was dependant on the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. WiDr cells had been seeded in 96-well plates at a thickness of just one 1 104 cells/well with 100 L of quantity and incubated at 37C and 5% CO2 for right away. WiDr cells had been added with several concentration from the nano-ASLE for 24 h. After that, the media had been removed. Next, it had HSL-IN-1 been added with 100 L of DMEM and 10 L MTT (5 mg MTT/mL alternative) to each and every well. The plates had been incubated for 4 h. Control cells received just the media with no nano-ASLE examples just simply. The crystal of formazan which shaped in practical cells was solubilized with 100 L of SDS-stopper HCl 0.1 N. The absorbance 595 nm was assessed by ELISA audience by Standard Microplate Audience (Bio-Rad, USA) and computed to look for the IC50 worth with linear regression evaluation using Microsoft Excel 2016. Immunocytochemistry staining WiDr cells had been seeded in 24-well plates at a thickness of.